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Cytotoxic Activities of Physalis minima L. Chloroform Extract on Human Lung Adenocarcinoma NCI-H23 Cell Lines by Induction of Apoptosis.

Leong OK, Muhammad TS, Sulaiman SF - Evid Based Complement Alternat Med (2011)

Bottom Line: Different stages of apoptotic programed cell death as well as phosphatidylserine externalization were confirmed using annexin V and propidium iodide staining.Furthermore, acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in this cell line.Due to its apoptotic effect on NCI-H23 cells, it is strongly suggested that the extract could be further developed as an anticancer drug.

View Article: PubMed Central - PubMed

Affiliation: School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.

ABSTRACT
Physalis minima L. is reputed for having anticancer property. In this study, the chloroform extract of this plant exhibited remarkable cytotoxic activities on NCI-H23 (human lung adenocarcinoma) cell line at dose- and time-dependent manners (after 24, 48 and 72 h of incubation). Analysis of cell-death mechanism demonstrated that the extract exerted apoptotic programed cell death in NCI-H23 cells with typical DNA fragmentation, which is a biochemical hallmark of apoptosis. Morphological observation using transmission electron microscope (TEM) also displayed apoptotic characteristics in the treated cells, including clumping and margination of chromatins, followed by convolution of the nuclear and budding of the cells to produce membrane-bound apoptotic bodies. Different stages of apoptotic programed cell death as well as phosphatidylserine externalization were confirmed using annexin V and propidium iodide staining. Furthermore, acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in this cell line. Due to its apoptotic effect on NCI-H23 cells, it is strongly suggested that the extract could be further developed as an anticancer drug.

No MeSH data available.


Related in: MedlinePlus

The effect of (a) DMSO l% (v/v), (b) Physalis minima chloroform extract (2.80 μg/mL, EC50 a 72 h), (c) DNase I (1 U/mL) and (d) vincrstine sulphate (0.0015 μg/mL, EC50 at 72 h) or NCI-H2: cells for 24 h and subjected to Deadend Colometric Apoptosis Detection System (Promega, USA) Apoptotic cells with stained nuclei were rnarked by arrows. (e) Comparison of the mean percentage of apoptotic index between DNase 1-, vincristine sulphate- and P. minima chloroform extract-treated cells to untreated cells (DMSO) at 24 hours treatment in different cell lines. Each value represented mean ± SEM from three independent experiments. *P < .05 (as compared with negative control).
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fig2: The effect of (a) DMSO l% (v/v), (b) Physalis minima chloroform extract (2.80 μg/mL, EC50 a 72 h), (c) DNase I (1 U/mL) and (d) vincrstine sulphate (0.0015 μg/mL, EC50 at 72 h) or NCI-H2: cells for 24 h and subjected to Deadend Colometric Apoptosis Detection System (Promega, USA) Apoptotic cells with stained nuclei were rnarked by arrows. (e) Comparison of the mean percentage of apoptotic index between DNase 1-, vincristine sulphate- and P. minima chloroform extract-treated cells to untreated cells (DMSO) at 24 hours treatment in different cell lines. Each value represented mean ± SEM from three independent experiments. *P < .05 (as compared with negative control).

Mentions: In order to investigate whether apoptosis may play an important role in mediating the cell death of NCl-H23 cells elicited by the chloroform extract of P. minima, the fragmented genomic DNA was detected using a modified TUNEL assay. As shown in Figures 2(b), 2(c) and 2(d), the extract-treated NCl-H23 cells produced dark brown stained nuclei with similar observation found in the positive control cells treated with DNase I and vincristine sulfate. The nuclei were specifically stained and evenly distributed. Most of the positive stained nuclei were rounded or oblong in shape. But almost all nuclei of untreated negative control cells were not stained with this assay (Figure 2(a)). The mean percentage of apoptotic index for the extract-treated NCl-H23 cells was 49.89% and significantly different as compared to the negative control (DMSO) (1.39%) (P < .05) (Figure 2(e). Comparatively, the percentage was lower than that in positive controls (68.41% and 56.40% for DNase I and vincristine sulfate, resp.). This result strongly indicated that apoptosis was one of the possible type of cell death in NCl-H23 cells after 24 h exposure to the chloroform extract. The trypan blue exclusion assay showed that only a few NCl-H23 cells were positively stained after treated with the extract for 24 and 72 h. The results strongly suggested that the surface of most treated cells was intact after incubation with the extract at both time points (Figures 3(a) and 3(b). Therefore, the mode of cell death elicited by the chloroform extract of P. minima in NCI-H23 cells was unlikely via necrotic mechanism in nature.


Cytotoxic Activities of Physalis minima L. Chloroform Extract on Human Lung Adenocarcinoma NCI-H23 Cell Lines by Induction of Apoptosis.

Leong OK, Muhammad TS, Sulaiman SF - Evid Based Complement Alternat Med (2011)

The effect of (a) DMSO l% (v/v), (b) Physalis minima chloroform extract (2.80 μg/mL, EC50 a 72 h), (c) DNase I (1 U/mL) and (d) vincrstine sulphate (0.0015 μg/mL, EC50 at 72 h) or NCI-H2: cells for 24 h and subjected to Deadend Colometric Apoptosis Detection System (Promega, USA) Apoptotic cells with stained nuclei were rnarked by arrows. (e) Comparison of the mean percentage of apoptotic index between DNase 1-, vincristine sulphate- and P. minima chloroform extract-treated cells to untreated cells (DMSO) at 24 hours treatment in different cell lines. Each value represented mean ± SEM from three independent experiments. *P < .05 (as compared with negative control).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3094824&req=5

fig2: The effect of (a) DMSO l% (v/v), (b) Physalis minima chloroform extract (2.80 μg/mL, EC50 a 72 h), (c) DNase I (1 U/mL) and (d) vincrstine sulphate (0.0015 μg/mL, EC50 at 72 h) or NCI-H2: cells for 24 h and subjected to Deadend Colometric Apoptosis Detection System (Promega, USA) Apoptotic cells with stained nuclei were rnarked by arrows. (e) Comparison of the mean percentage of apoptotic index between DNase 1-, vincristine sulphate- and P. minima chloroform extract-treated cells to untreated cells (DMSO) at 24 hours treatment in different cell lines. Each value represented mean ± SEM from three independent experiments. *P < .05 (as compared with negative control).
Mentions: In order to investigate whether apoptosis may play an important role in mediating the cell death of NCl-H23 cells elicited by the chloroform extract of P. minima, the fragmented genomic DNA was detected using a modified TUNEL assay. As shown in Figures 2(b), 2(c) and 2(d), the extract-treated NCl-H23 cells produced dark brown stained nuclei with similar observation found in the positive control cells treated with DNase I and vincristine sulfate. The nuclei were specifically stained and evenly distributed. Most of the positive stained nuclei were rounded or oblong in shape. But almost all nuclei of untreated negative control cells were not stained with this assay (Figure 2(a)). The mean percentage of apoptotic index for the extract-treated NCl-H23 cells was 49.89% and significantly different as compared to the negative control (DMSO) (1.39%) (P < .05) (Figure 2(e). Comparatively, the percentage was lower than that in positive controls (68.41% and 56.40% for DNase I and vincristine sulfate, resp.). This result strongly indicated that apoptosis was one of the possible type of cell death in NCl-H23 cells after 24 h exposure to the chloroform extract. The trypan blue exclusion assay showed that only a few NCl-H23 cells were positively stained after treated with the extract for 24 and 72 h. The results strongly suggested that the surface of most treated cells was intact after incubation with the extract at both time points (Figures 3(a) and 3(b). Therefore, the mode of cell death elicited by the chloroform extract of P. minima in NCI-H23 cells was unlikely via necrotic mechanism in nature.

Bottom Line: Different stages of apoptotic programed cell death as well as phosphatidylserine externalization were confirmed using annexin V and propidium iodide staining.Furthermore, acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in this cell line.Due to its apoptotic effect on NCI-H23 cells, it is strongly suggested that the extract could be further developed as an anticancer drug.

View Article: PubMed Central - PubMed

Affiliation: School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.

ABSTRACT
Physalis minima L. is reputed for having anticancer property. In this study, the chloroform extract of this plant exhibited remarkable cytotoxic activities on NCI-H23 (human lung adenocarcinoma) cell line at dose- and time-dependent manners (after 24, 48 and 72 h of incubation). Analysis of cell-death mechanism demonstrated that the extract exerted apoptotic programed cell death in NCI-H23 cells with typical DNA fragmentation, which is a biochemical hallmark of apoptosis. Morphological observation using transmission electron microscope (TEM) also displayed apoptotic characteristics in the treated cells, including clumping and margination of chromatins, followed by convolution of the nuclear and budding of the cells to produce membrane-bound apoptotic bodies. Different stages of apoptotic programed cell death as well as phosphatidylserine externalization were confirmed using annexin V and propidium iodide staining. Furthermore, acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in this cell line. Due to its apoptotic effect on NCI-H23 cells, it is strongly suggested that the extract could be further developed as an anticancer drug.

No MeSH data available.


Related in: MedlinePlus